towards gene therapy for duchenne muscular dystro

... PROGENITOR CELLS AS TARGETS FOR GENE THERAPY 143 144 GENE THERAPY FOR HEMATOLOGICAL DISORDERS restoration of cellular and humoral immunity were documented after gene therapy Data showed a surprisingly ... otherwise standard MuLV vectors for gene therapy has not been successful Beyond these HEMATOPOIETIC STEM AND PROGENITOR CELLS AS TARGETS FOR GENE THERAPY 137 efforts, there are obviously major ... In vivo gene transfer is most appropriate for target cells that cannot REQUIREMENTS FOR GENE TRANSFER INTO HEMATOPOIETIC CELLS 135 TABLE 6.1 Relevant Targets and Applications for Gene Therapy...

Ngày tải lên: 10/04/2014, 22:10

... hand, it is necessary for any vehicle used for in 156 GENE THERAPY FOR LIVER DISEASE vivo hepatic gene therapy to reach the liver efficiently For systemic application, the gene vectors are ideally ... attempted so far GENERAL PRINCIPLES FOR HEPATIC GENE THERAPY There are two basic approaches for gene transfer into hepatocytes: ex vivo and in vivo strategies (Fig 7.1) Ex vivo therapy requires ... expression of the therapeutic gene. Alternatively, different viral vectors as well as liposomes can be used for ex vivo gene transfer GENERAL PRINCIPLES FOR HEPATIC GENE THERAPY 155 Collagenase treatment...

Ngày tải lên: 10/04/2014, 22:11

... MKL, Morgan RA Gene therapy for acquired immune deficiency syndrome AIDS Updates 7:1–14, 1994 Yu M, Poeschla E, Wong-Staal F Progress towards gene therapy for HIV infection Gene Therapy 1:13–26, ... gene expression Thus, the antisense gene expression must be quantatively higher than the levels of HIV-1 gene expression for an antisense gene therapy strategy to be effective Standard gene therapy ... levels in HSCs Thus, retroviral-mediated gene transfer is currently the optimal gene transfer system available for use in HIV-1 gene therapy The 282 GENE THERAPY FOR HIV INFECTION limitations of retroviral...

Ngày tải lên: 10/04/2014, 22:11

... performed twice postbone marrow transplantation (BMT) at days 730 and 1250 to study histopathology, dystrophin expression and chimerism status Immunostaining for dystrophin I was reduced, dystrophin ... parameters were optimized as follows: 95°C for 11 minutes (one hold), 94°C for 60 seconds, 59°C for 60 seconds, 72°C for 60 seconds and 28 cycles, 60°C for Figure Hematoxylin and eosin-stained ... noted 60 minutes (two holds) in a GeneAmp PCR System 9700 (Applied Biosystems) Denaturation was performed for five minutes at 95°C using Hi-Di Formamide and GeneScan 500 LIZ Size Standard (both...

Ngày tải lên: 10/08/2014, 23:21

... to be answered for successful CF gene therapy have now been clearly defined, and gene therapy for CF remains the most promising possibility for curative rather than symptomatic therapy 12 13 14 ... models that successful gene therapy for cystic fibrosis requires only partial gene correction Gene Ther 1996, 3:797–801 28 Hyde SC, Southern KW, Gileadi U, Fitzjohn EM, Mofford KA, Waddell BE, ... has dogged the recent media view of gene therapy Gene therapy is clearly at the typical stage of new drug development where considerable and unglamorous effort needs to be expended to move from...

Ngày tải lên: 12/08/2014, 18:20

... Therefore, in the particular case of Epo, an accurate level of transgene expression is required for safety reasons Temporal control systems of transgene expression have already been used in gene therapy ... erythropoietin gene therapy for patients with chronic renal failure Blood 2005, 106(7):2280-2286 Osada S, Ebihara I, Setoguchi Y, Takahashi H, Tomino Y, Koide H: Gene therapy for renal anemia ... using plate electrodes and generator BTX ECM 830 (Genetronics™), as previously described [20] Page of (page number not for citation purposes) Genetic Vaccines and Therapy 2008, 6:10 Statistical...

Ngày tải lên: 14/08/2014, 19:22

... with an exposure time of 50 ms for DAPI and 500 ms for EGFP (1 s for negative expression) Page of (page number not for citation purposes) Genetic Vaccines and Therapy 2009, 7:9 http://www.gvt-journal.com/content/7/1/9 ... purposes) Genetic Vaccines and Therapy 2009, 7:9 Acknowledgements http://www.gvt-journal.com/content/7/1/9 18 We thank Günter Schwarz (Köln) for providing cPMP and Sebastian Kügler (Göttingen) for ... borderline result and indicates a range for the minimal dosage required for abolishing the MoCo deficiency phenotype With a maximum body weight of 40 g for the mice used here this would correspond...

Ngày tải lên: 14/08/2014, 19:22

... When dystrophin is absent, the DGC is destabilized leads to muscle wasting which characterizes Duchenne muscular dystrophy 1.4 Mutations in the dystrophin gene Mutations in the large dystrophin gene, ... of different isoforms (Figure 3) [35] 10 Figure 3: A: location of DMD gene B: protein products of the DMD gene [35] 1.3 Protein dystrophin The product of the dystrophin gene is dystrophin protein ... Amplification” CHAPTER INTRODUCTION 1.1 Duchenne muscular dystrophy 1.1.1 Characteristics of DMD Duchenne muscular dystrophy (DMD) was first described by Duchenne de Boulogne, the French neurologist,...

Ngày tải lên: 05/11/2015, 17:38

... detection in families affected by Duchenne muscular dystrophy using Multiplex Ligation- dependent Probe Amplification” CHAPTER INTRODUCTION 1.1 Duchenne muscular dystrophy 1.1.1 Characteristics ... 116, Dp 71, leading to a number of different isoforms (Figure 3) [35] 1.3 Protein dystrophin The product of the dystrophin gene is dystrophin protein Dystrophin is a hydrophobic, rod-shaped protein ... laboratories performing genetic testing for the molecular diagnosis in general and DMD in particular Compared to other techniques, MLPA is rapid, sensitive, reliable and very simple to perform Therefore,...

Ngày tải lên: 05/11/2015, 17:39

... vectors for gene therapy formed by self-assembly of DNA with synthetic block co-polymers Hum Gene Ther 7, 2123–2133 11 Kabanov, A V and Kabanov, V A (1995) DNA complexes with polycations for the ... refs 8–10 for typical results) The protocol can be used for all adherent cell types For suspension type cells, cells need to be spun down before changing the medium at a speed suitable for the ... tetrazolium salt, XTT, to form an orange formazan dye by dehydrogenase activity in active mitochondria Therefore, this conversion only occurs in living cells The formazan dye formed is soluble in...

Ngày tải lên: 11/04/2014, 09:59

... neurodegeneration (133) Dystrophin in Duchenne muscular dystrophy (95) Anti-inflammatory cytokines for arthritis (134,135) A variety of immunomodulatory, suicide, and radiotherapy-enhancing genes ... vectors for gene transfer and gene therapy I would like to thank the many chapter authors for their contributions They are all experts in various aspects of viral vectors, and I appreciate their efforts ... Type 1-Based Vectors for Gene Delivery to Human Hematopoietic Stem Cells Ali Ramezani and Robert G Hawley 467 Semliki Forest Viral Vectors for Gene Transfer Jarmo Wahlfors and Richard A Morgan...

Ngày tải lên: 11/04/2014, 10:39

... important for the resistance to possible introduction of genes using viral vectors or for the resistance to oncolytic gene therapy We believe that this finding can be of crucial interest for the ... viral gene therapy approaches Adenoviruses and Adeno-Associated viruses are used to deliver genes to tumor cells with the goal of modifying the phenotype, as for example, by introducing suicide genes ... primers: Ad type forward primer 5’-AACCGAAGGCTGCATTCACT, reverse primer 5’-ACCGCACAGGGTCTTAATAGAG Following denaturation at 96°C for 10 min, cycling conditions were 96°C for 15s, 60°C for for 40 cycles...

Ngày tải lên: 18/06/2014, 16:20

... as follows: first cycle at 95°C for min, and then 30 cycles at 94°C for 45 s, 54°C for min, 72°C for and a final extension cycle of 72°C for The house-keeping gene b-actin was taken as a loading ... DNA for gene therapy [9,14,15] For example, a liner PEI-cholesterol conjugation was encapsulated with interleukin-12 to treat RCC mice intravenously, which was demonstrated to be effective for ... weight, and the gene therapy effects on RCC model mice were obvious by using FA-PEAs:pVHL complexes to treat tumor Therefore, FA-PEAs may be a potential gene transfer system to carry VHL gene to treat...

Ngày tải lên: 18/06/2014, 19:20

... as follows: first cycle at 95°C for min, and then 30 cycles at 94°C for 45 s, 54°C for min, 72°C for and a final extension cycle of 72°C for The house-keeping gene b-actin was taken as a loading ... DNA for gene therapy [9,14,15] For example, a liner PEI-cholesterol conjugation was encapsulated with interleukin-12 to treat RCC mice intravenously, which was demonstrated to be effective for ... weight, and the gene therapy effects on RCC model mice were obvious by using FA-PEAs:pVHL complexes to treat tumor Therefore, FA-PEAs may be a potential gene transfer system to carry VHL gene to treat...

Ngày tải lên: 20/06/2014, 03:20

... and thus the former is the vector of choice for future gene therapy trials The number of gene therapy trials in RA is in sharp contrast to the more than 600 clinical trials of gene therapy in patients ... safety of local gene therapy Gene therapy in RA is still in its adolescence, and more clinical studies must be initiated if it is to mature It is, however, imperative that gene therapy studies ... Sacchi A: Wild-type p53 gene transfer is not detrimental to normal cells in vivo: implications for tumor gene therapy Oncogene 2004, 23:418-425 Kruyt FA, Curiel DT: Toward a new generation of conditionally...

Ngày tải lên: 09/08/2014, 01:24

... Thus, CD34 progenitor cells are an essential ingredient for HIV gene therapy In view of the need for CD34 cells for HIV gene therapy as well as for other hematopoietic disorders, if one can produce ... lineages thus paving the way for utilization of these cells for hematopoietic cell therapy [20,27-29] For the effective utilization of hES-CD34 cells for HIV gene therapy, a number of parameters ... Stem Cell-based Gene Therapy for HIV/AIDS Anticancer Res 2003, 23:1997-2005 Michienzi A, Castanotto D, Lee N, Li S, Zaia JA, Rossi JJ: RNA-mediated inhibition of HIV in a gene therapy setting...

Ngày tải lên: 13/08/2014, 09:21

... number not for citation purposes) Retrovirology 2004, 1:29 40 41 42 http://www.retrovirology.com/content/1/1/29 Yu M, Poeschla E, Wong-Staal F: Progress towards gene therapy for HIV infection Gene ... observed, and these macaques maintained high CD4+ T-lymphocyte counts for at least 478 days [24] However, a gene therapy strategy for HIV infection would only be possible during the chronic phase ... low-level autocrine IFN-β production as an approach to gene therapy for AIDS The persistence of transduced cell per 103 circulating cells before SIV challenge was correlated with low plasma virus...

Ngày tải lên: 13/08/2014, 13:20

... http://www.ncbi.nlm.nih.gov/nuccore/9629357?report=fasta General discussion I report here SIV/HIV-pol gene and HIV-1 whole genome specific zinc finger nucleases, which are proposed for use towards targeted HIV gene therapy Specifically, ... Annu Rev Pharmacol Toxicol 2011, 51:397-418 12 Wayengera M: HIV and Gene Therapy: The proposed [R-M enzymatic] model for a gene therapy against HIV Makerere Med J 2003, 38:28-30 13 Wayengera M, ... Microbicides for preventing HIV infection among high-risk women Afr J Biotechnol 2007, 6(10):1221-1224 18 Wayengera M: Pre-Integration gene slicing (PRINT-GSX) as an alternate or complimentary gene therapy...

Ngày tải lên: 13/08/2014, 16:20

... tomography of vector-mediated gene expression in gene therapy for gliomas Lancet 2001, 358:727-729 Page 12 of 13 (page number not for citation purposes) Genetic Vaccines and Therapy 2004, 2:7 http://www.gvt-journal.com/content/2/1/7 ... transduction levels, transgene localization and time course of fusion gene expression in living animals by BLI can aid in developing more potent gene therapy vectors for treatment of malignant ... at generating a sensitive tool for noninvasive in vivo monitoring of the activity of a therapeutic transgene by fusing the bioluminescent reporter gene Luc to the bioactivating "suicide" gene...

Ngày tải lên: 14/08/2014, 19:22

... retroviral genes Oncogene capture The mechanism of oncogene capture appears to be dependent on the generation of a chimeric retroviraloncogene transcript (69, 70) This suggests that the risk of oncogene ... or necessity? Insertional mutagenesis in gene therapy and its consequences Mol Ther 2004, 9:5-13 Dave UP, Jenkins NA, Copeland NG: Gene therapy insertional mutagenesis insights Science 2004, ... Page of 13 (page number not for citation purposes) Genetic Vaccines and Therapy 2004, 2:9 promoter used to drive transcription of the transgene (see below) Retroviral gene transfer The minimization...

Ngày tải lên: 14/08/2014, 19:22